Page 45 - Book of Abstracts

5 International Scientific Online Conference DOI: https://doi.org/10.15414/2021.9788055224015

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INFLUENCE OF CONCENTRATION OF AQUEOUS-ETHANOL SOLUTION ON EXTRACTION
OF FLAVONOIDS FROM "HAIRY" ROOTS OF MEDICINAL PLANTS
Volodymyr Duplij, Nadiia Matvieieva
Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences
of Ukraine, Kyiv, Ukraine; E-mail.: joyna56@gmail.com
Plants have been used by mankind since ancient times for the treatment and
prevention of various diseases. However, mass collection of plant raw materials can pose a
threat to natural populations. In addition, contamination of the environment with toxicants
sometimes makes it impossible to collect plants that grow naturally. The use of
biotechnological approaches, in particular, Agrobacterium-mediated transformation, allows
obtaining "hairy" roots of medicinal plants. Such roots are a promising source of a complex
of bioactive compounds, as they can synthesize compounds inherent in plants, and their
number may be many times higher than that in control plants. In addition, the roots grown
in bioreactors are not exposed to negative natural factors that reduce the content of valuable
compounds, in particular, flavonoids, as they are cultivated under standardized conditions
(medium composition, temperature, humidity, etc.). However, the extraction conditions of
the compounds synthesized in "hairy" roots are an important technological factor that must
be taken into account. In this work, we analyzed the features of extraction of flavonoids
using a water-ethanol mixture with different ethanol content. "Hairy" roots of Cichorium
intybus L. (2 samples) and Artemisia vulgaris L. (2 samples) were grown under standard
conditions (Murashige and Skoog nutrient medium), collected, lyophilized, and crushed.
Extraction was performed at + 25 °C for 24 h using 40 %, 70 % and 96 % aqueous-ethanol
mixture. The total flavonoid content was determined by reaction with aluminum chloride
per 1 gram of dry weight (DW).
It was found that the studied root samples differed significantly in the content of
flavonoids after extraction with a water-ethanol mixture of different concentrations. Thus,
in the A. vulgaris sample №1 the largest amount of flavonoids was detected by extraction
with 40 % ethanol – 63.5 ±1.25 mg/g DW. When 70 and 96 % ethanol were used, the amount
of flavonoids was 25.2 ±0.28 and 35.18 ±2.15 mg/g DW, respectively. At the same time, the
detected content of flavonoids in sample №2 of the same species was 55.49 ±3.34, 67.78
±7.03, and 73.08 ±4.28, respectively, using 40 %, 70 %, and 96 % ethanol.
Two samples of C. intybus differed from the above-described extracts because a higher
content of flavonoids was detected using 70 % ethanol. Thus, flavonoid content was 74.82
±1.56, 89.35 ±1.32, and 84.48 ±3.12 in the line №1, as well as in the line №2 it was 71.34
±1.79, 93.99 ±1.51, and 88.34 ±1.31 mg/g DW when used respectively 40 %, 70 %, and 96
% ethanol.
Such features can be explained by the different qualitative compositions of flavonoids.
This is due to the solubility of glycosylated flavonoids (preferably extracted with a water-
ethanol mixture with a lower ethanol concentration) and aglycones, which are extracted with a
mixture with a high concentration of ethanol. The obtained data indicate a probably higher
content of aglycones in two samples of chicory "hairy” roots in comparison with the wormwood
sample №1. Thus, when optimizing the method of extraction of flavonoids, it is necessary to
conduct preliminary testing using an aqueous-ethanol mixture of different concentrations for
maximum extraction f the target compounds.

Keywords: Cichorium intybus, Artemisia vulgaris, "hairy" roots, flavonoids.
Acknowledgments
The work was supported by a grant № II-2-21of the National Academy of Sciences of Ukraine.

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